Raman Spectroscopy Detects Biochemical Changes Due to Different Cell Culture Environments in Live Cells In Vitro

The in vitro cell culture environment can impact on cell biochemistry and cell cycle. The manifestation of such substrate-induced changes in cell cycle in the Raman microspectroscopic profiles of cell cultures is investigated at the level of nucleolus, nucleus and cytoplasm. HeLa immortalised human cervical cells and HaCaT dermal cells were cultured on three different substrates, conventional polystyrene cell culture dishes, CaF2 slides as a commonly used Raman substrate, and glass slides coated with collagen rat tail, as a mimic of the extra-cellular matrix (ECM) environment. A cell cycle study, based on percentage DNA content, as determined using propidium iodide staining and monitored by flow cytometry, was performed on cells of both types, grown on the different substrates, confirming that the in vitro cell culture environment impacts significantly on the cell cycle. Live cell in vitro Raman spectroscopic analysis of cells on the 2D CaF2 and 3D collagen substrates was performed and data was analysed using principal component analysis (PCA). The spectroscopic analysis revealed differences in profiles which reflect the differences in cell cycle for both in vitro culture environments. In particular, the Raman spectra of cells grown on CaF2 show indicators of cell stress, which are also associated with cell cycle arrest at the G0/G1 phase. This work contributes to the field of Raman spectroscopic analysis by providing a fresh look at the significance of the effect of in vitro culture environment to cell cycle and the sensitivity of Raman spectroscopy to such differences in cell metabolism

Comparison of time-gated surface-enhanced raman spectroscopy (TG-SERS) and classical SERS based monitoring of Escherichia coli cultivation samples

The application of Raman spectroscopy as a monitoring technique for bioprocesses is severely limited by a large background signal originating from fluorescing compounds in the culture media. Here, we compare time-gated Raman (TG-Raman)-, continuous wave NIR-process Raman (NIR-Raman), and continuous wave micro-Raman (micro-Raman) approaches in combination with surface enhanced Raman spectroscopy (SERS) for their potential to overcome this limit. For that purpose, we monitored metabolite concentrations of Escherichia coli bioreactor cultivations in cell-free supernatant samples. We investigated concentration transients of glucose, acetate, AMP, and cAMP at alternating substrate availability, from deficiency to excess. Raman and SERS signals were compared to off-line metabolite analysis of carbohydrates, carboxylic acids, and nucleotides. Results demonstrate that SERS, in almost all cases, led to a higher number of identifiable signals and better resolved spectra. Spectra derived from the TG-Raman were comparable to those of micro-Raman resulting in well-discernable Raman peaks, which allowed for the identification of a higher number of compounds. In contrast, NIR-Raman provided a superior performance for the quantitative evaluation of analytes, both with and without SERS nanoparticles when using multivariate data analysis. (c) 2018 American Institute of Chemical EngineersPeer reviewe

Hybrid EEFIT mission to february 2023 Kahramanmaraş earthquake sequence

The southwestern part of Türkiye was hit on 6 February 2023 by an Mw 7.8 (epicentre:Pazarcık) and then an Mw 7.5 earthquake (epicentre: Elbistan). The event was followed by tensof thousands of aftershocks including the Mw 6.3 event on 20 February (epicentre: Uzunbağ).This paper reports on the preliminary findings of the mission organised by the UK’s EarthquakeEngineering Field Investigation Team (EEFIT) to the Kahramanmaraş Earthquake sequence ofFebruary 2023. This mission followed a hybrid model, combining field and remote investigationtechniques, to investigate the characteristics of the earthquake sequence, its impact on buildingsand infrastructure, as well as the efficacy of relief, response and recovery operations. The keymessages include that the building stock is hard to categorise which brings along difficulties withdamage assessment, that the recovery and reconstruction require multi-sectoral engagement ofkey stakeholders, and that the auditing and quality control mechanisms within the constructionindustry need revisiting in the way forward for better disaster resilience in Türkiye

The Türki̇ye earthquake sequence of February 2023: A longitudinal study report by EEFIT

On 6 February 2023 at 4:17 am local time, a large area in southeastern Türkiye and northern Syria was hit by an Mw 7.8 earthquake, which was followed by an Mw 7.5 earthquake at 1:24 pm local time, causing the loss of more than 50,000 lives, some 100,000 injuries and significant damage to buildings and infrastructure, estimated to be in the range of 84.1 billion USD for Türkiye alone. The largest earthquake in Türkiye since the deadly 1939 Erzincan earthquake with however much larger losses, the sequence immediately attracted the attention of the global post-disaster reconnaissance/engineering communities. This included the Earthquake Engineering Field Investigation Team (EEFIT), who, within one week of the event, gathered a team with 30 people from academia and industry in the UK (19), Türkiye (5), New Zealand (1), Hungary (1), Bulgaria (1), Greece (1) and USA (1) with two support members from the UK and the Netherlands, to study the events and their impacts, and also to develop suggestions to reduce the existing vulnerabilities in the future. The team was organised in the form of 6 working groups as shown below, which were (1) strong ground motions and seismotectonics, (2) geotechnics, (3) structures, (4) infrastructure, (5) remote sensing and (6) relief response and recovery

2023 Türki̇ye deprem sekansi deprem mühendi̇sli̇ği̇ saha çalişmalari eki̇bi̇ (EEFIT) Misyonu: Türkçe geni̇şleti̇lmi̇ş özet

Türkiye'nin güneydoğusunda ve kuzey Suriye'de geniş bir alan, 6 Şubat 2023 tarihinde yerel saatle sabaha karşı 04:17'de gerçekleşen 7.8 büyüklüğünde yıkıcı bir depremle sarsıldı, aynı gün içerisinde 13:24’te meydana gelen 7.5 büyüklüğündeki deprem ile yıkımın boyutu ve etki alanı büyük ölçüde arttı. Bu depremler, yalnızca Türkiye'de 84.1 milyar $’ı (≈2.5 trilyon ₺) aşan ekonomik kayba, bina ve altyapı stokunda ciddi hasara, 100,000'den fazla yaralı ve 50,000'den fazla can kaybına neden oldu

Method to obtain platelet-rich plasma from rabbits (Oryctolagus cuniculus )

Abstract: Platelet-rich plasma (PRP) is a product easy and inxpesnsive, and stands out to for its growth factors in tissue repair. To obtain PRP, centrifugation of whole blood is made with specific time and gravitational forces. Thus, the present work aimed to study a method of double centrifugation to obtain PRP in order to evaluate the effective increase of platelet concentration in the final product, the preparation of PRP gel, and to optimize preparation time of the final sample. Fifteen female White New Zealand rabbits underwent blood sampling for the preparation of PRP. Samples were separated in two sterile tubes containing sodium citrate. Tubes were submitted to the double centrifugation protocol, with lid closed and 1600 revolutions per minute (rpm) for 10 minutes, resulting in the separation of red blood cells, plasma with platelets and leucocytes. After were opened and plasma was pipetted and transferred into another sterile tube. Plasma was centrifuged again at 2000rpm for 10 minutes; as a result it was split into two parts: on the top, consisting of platelet-poor plasma (PPP) and at the bottom of the platelet button. Part of the PPP was discarded so that only 1ml remained in the tube along with the platelet button. This material was gently agitated to promote platelets resuspension and activated when added 0.3ml of calcium gluconate, resulting in PRP gel. Double centrifugation protocol was able to make platelet concentration 3 times higher in relation to the initial blood sample. The volume of calcium gluconate used for platelet activation was 0.3ml, and was sufficient to coagulate the sample. Coagulation time ranged from 8 to 20 minutes, with an average of 17.6 minutes. Therefore, time of blood centrifugation until to obtain PRP gel took only 40 minutes. It was concluded that PRP was successfully obtained by double centrifugation protocol, which is able to increase the platelet concentration in the sample compared with whole blood, allowing its use in surgical procedures. Furthermore, the preparation time is appropriate to obtain PRP in just 40 minutes, and calcium gluconate is able to promote the activation of platelets

Papillon-Lefevre syndrome - Analysis of peripheral blood lymphocyte subsets

We have studied the peripheral blood lymphocyte populations in our 6 patients (2 female and 4 male) with a mean age of 11.19 with Papillon-Lefevre Syndrome (PLS) using adequate monoclonal antibodies and double coloured flow cytometry. Total B, T, CD4, CD8, CD29, CD45RA, NK, HLA-DR cells were studied. Total B, T, CD4 and CD8 lymphocytes were within normal limits. We have observed an increase in the CD29 lymphocytes and NK cells and a decrease in CD45RA lymphocytes. We think that these findings are important in explaining B lymphocyte activation and in the pathogenesis of the PLS